Microfluidic-based 3d fibroblast migration studies in biomimetic microenvironments

Abstract

Cell migration in 3D is a fundamental process in many physiological and pathological phenomena. Indeed, migration through interstitial tissue is a multi-step process that turns out from the cell-ECM interaction. It is a dynamic and complex mechanism that depends on the physic-chemical balance between the cell and its surrounding. Early stage of deep dermal wound healing process is a relevant migratory example, in which the fibroblast is the epicenter: the recruitment of the fibroblasts -by chemotaxis of PDGF-BB- to the clotted wound occurs. Likewise, this work focuses on studying the major underlying mechanisms of 3D fibroblast migration and the main microenvironmental cues involved within. To do so, we have confined two physiologically relevant hydrogels, made of collagen and fibrin, within microfluidic platforms. Firstly, an integral comparative study of biophysical and biomechanical properties of both gels is presented. In these results, we have overcome the wide diversity of the existing data and special stress has been done in order to compare the microstructural arrangement, resistance to flow and elasticity. On the other hand, controlled chemical gradients have been generated and characterized within the microfluidic devices. Since biomolecules interact as purely diffusive factors or bound to the matrix proteins, in this work, distribution of PDGF-BB and TGF-ß1 across collagen and fibrin gels has been quantified. Finally, by taking advantage of the biophysico-chemical definition, we have characterized the migratory responses of human fibroblasts within the microsystems in the presence of a chemoattractant (PDGF-BB). Our results demonstrate that the local microarchitecture of the hydrogels determines the migratory properties of human fibroblasts in response to controlled chemotactic and haptotactic gradients, in a myosin II-dependent manner.